UCLA researchers in the Department of Molecular, Cell, and Developmental Biology have developed a novel high-throughput method for the quantification of immune cell subtype.
Quantification of leukocytes (white blood cells) among peripheral blood mononuclear cells (PBMCs) may provide insights to the immune-biology of auto-immune disorders and infectious diseases. Flow cytometry is the current standard for clinical assessment but it is expensive, limited to fresh samples, and has limited throughput. DNA methylation plays important roles in development and cellular differentiation. Previous work has demonstrated that differentiated cells and tissues have unique DNA methylation patterns. New approaches that measure DNA methylation using sequencing instead of probe-based arrays provide the advantage of single base-pair resolution, and the potential for 10-100 times more cytosines methylation status data. However, a robust high-throughput method using this methylation data to identify immune cell subtypes has yet to be developed.
UCLA researcher Dennis Montoya and Prof. Matteo Pellegrini and colleagues have developed a high-throughput method that utilizes bisulfite sequencing to yield a robust platform for decomposing blood profiles into their constituent cell types. Through the selection of cell type specific DNA methylation signatures and linear regression, the technology can reconstruct cell type proportions from in vitro cell mixture experiments. This method provides an approach to quantify cell type quantities with greater power over DNA methylation array-based approaches with potential applications beyond immune-biology.
This innovation has been validated in vitro and work is underway to validate testing of clinical PBMCs.
High-throughput sequencing, CpG methylation, cytosine methylation, DNA methylation, immune cells, white blood cells, PBMCs, peripheral blood mononuclear cells, cell quantification, microarray, leukocytes